Membrane deletion during plasmolysis in hardened and non-hardened plant cells

Abstract Video recordings of interference phase contrast microscopy were used to study plasmalemma deletion during plasmolysis in hardened and non-hardened suspension cultured cells of Brassica napus, alfalfa, and cells isolated from rye seedlings. Although different hardening regimes and different cells were used, the responses to plasmolysis were consistent. Hardened cells uncoupled the volume to surface area ratio during plasmolysis both by forming a large number of strands between the cell wall and protoplast and by leaving rivulet-like networks of membranes on the cell wall surface. Tonoplast membrane was deleted as sac-like intrusions into the vacuole. Non-hardened cells produced few strands during plasmolysis. They also deleted plasmalemma and tonoplast into the vacuole as endocytotic vesicles. During deplasmolysis of hardened cells both the individual membrane strands and the rivulets of membrane material vesiculated into strings of vesicles. The vesicles were osmotically active and were re-incorporated into the expanding protoplast. Conversely, deplasmolysis in non-hardened cells resulted in few osmotically active vesicles and many broken strands. The vacuolar sac-like intrusions in hardened cells were re-incorporated into the vacuole whereas the endocytotic vesicles in non-hardened cells were not re-incorporated. Therefore, the non-hardened cells underwent expansion-induced lysis.     

Photosynthetic pathway,chilling tolerance and cell sap osmotic potential values of grasses along an altitudinal gradient in Papua New Guinea

Summary A total of 22 grass species were examined from 5 sites spanning the altitudinal range 1550–4350 m.a.s.l. The presence of the C₃ or C₄ photosynthetic pathway was determined from ¹³C values and chilling tolerance was assessed on the basis of electrolyte leakage from leaf slices incubated on melting ice. Most of the grasses studied at the lower altitude sites of 1550 m.a.s.l. (annual mean of daily minimum temperature, 14.6° C) and 2600 m.a.s.l. (9.4° C) possessed C₄ photosynthesis and were chill-sensitive. The single except ion was Agrostis avenacea, a montane chill-resistant C₃ species which occurred at 2600 m.a.s.l. The three species apparently most sensitive to chilling were Ischaemum polystachyum, Paspalum conjugatum and Saccharum robustum, all occurring at 1550 m.a.s.l. At the higher altitude sites of 3280 (5.6° C), 3580 (4.0° C) and 4350 (–0.7°C) m.a.s.l., most of the grasses exhibited C₃ photosynthesis and were chill-resistant. However, an Upland population of the C₄ species, Miscanthus floridulus was found at 3280 m.a.s.l. which had acquired chill-resistance as confirmed by additional in vivo variable chlorophyll fluorescence measurements. Cell sap osmotic potential values of the upland grasses at altitudes of 3280–4350 m.a.s.l. were lower (–8.1 to –19.8 bars) than values in grasses from 1550 and 2600 m.a.s.l. (–3.9 to –7.5 bars) due mainly to the presence of non-electrolyte osmoticants, which may be involved in frost avoidance mechanism(s).Abbreviations ABA abscisic acid - F_R the maximal rate of rise of induced chlorophyll fluorescence - s osmotic potential     

Accelerated nitrogen inputs — A new problem or a new perspective?

This paper considers whether new problems are arising in forest ecosystems due to increased levels of inorganic N deposition from the atmosphere, or whether there are no new problems, just a change of perception. Deposition of N has indeed increased. Wet deposition rates are reasonably quantified, but the rates of dry deposition to forests are largely unknown. Current transport and deposition models are probably under-estimating N deposition to forests. Consideration of possible effects of enhanced N deposition reveals with varying degrees of certainty that there may be effects due to high N in biomass, high uptake rates, leaching of nitrate and consequent acidification, and an overall increase in N availability. Forest ecosystems are not well-enough understood to set a critical load for N deposition, but enough is known to define some upper limits.     

Descending influences from nucleus raphe magnus on fusimotor neurone activity in rats

1. 1.Single fusimotor fibres were isolated in the ventral roots of lumbosacral segments of urethane-anaesthetized rats, and effects of electrical stimulation of the nucleus raphe magnus (NRM) on their spontaneous activity were investigated. The experiments were carried out in rats whose bilateral preoptic and anterior hypothalamic regions (PO/AH) were electrolytically destroyed to eliminate the influences of these regions to fusimotor activity.

2. 2.Of 44 fusimotor fibres studied, 38 (86%) were found to be affected by NRM stimulation. The effects of NRM stimulation were classified according to their response pattern: primary depression (D-type, n = 24), facilitation followed by depression (F-D-type, n = 5) and primary facilitation (F-type, n = 8). The most predominant effect of NRM stimulation upon fusimotor activity was characterized by a strong depression followed by a complete cessation of firing lasting either for a short period or for more than 30 min (D- and F-D-type).

3. 3.In three fusimotor fibres studied in the different preparations, it was observed that a NRM-evoked depression response was blocked by an intraperitoneal administration of a serotonin antagonist, p-chlorophenylalamine (p-CPA) (10 mg/kg).

4. 4.The results indicate that the NRM exerts descending inhibitory or facilitatory influences on fusimotor neurones, and suggest that cold shivering is controlled by modulating fusimotor neurone activity via the serotonergic raphe-signal pathways.

Participation of the cyanide-resistant pathway in respiration of winter rape leaves as affected by plant cold acclimation

Leaf slices sampled from winter rape plants ( Brassica napus L., var. oleifera L., cv. GórczaánAski), grown in cold (5°C), showed an increase in the dark respiration rate (measured at 25°C) as compared to slices cut from control plants (grown at 20/15°C). The effect of low temperature was most pronounced after 4 days of plant growth in the cold. Oxygen uptake by control slices was 60% inhibited by 1 m M KCN and was insensitive to 2.5 m M salicylhydroxamic acid (SHAM). On the contrary, respiration of leaf slices from cold-pretreated plants was more resistant to cyanide (35% inhibition after 4 days of cold treatment) and was 30% inhibited by SHAM. The patterns of cold-induced changes in total respiratory activity and in the estimated activity of alternative pathway were similar. It seems that in leaf slices from plants grown in the cold, the cyanide-resistant, alternative pathway participates in oxygen uptake. Cold treatment of plants also brought about a 4-fold increase in the level of soluble sugars, which reached a maximum on day 4 of exposure to cold. Addition of sucrose to the incubation medium resulted in an immediate increase in oxygen uptake by slices with low endogenous sugar level. The respiration stimulated by sucrose addition was more resistant to cyanide than the basal respiration and it was inhibited by SHAM. It is concluded that the operation of the alternative pathway is responsible for the increased oxygen uptake by the cold-grown winter rape leaves and it may be induced by an increased sugar supply for respiratory processes.     

Monitoring chilling injury: A comparison of chlorophyll fluorescence measurements, post-chilling growth and visible symptoms of injury in Zea mays

Changes in chlorophyll fluorescence emission from maize ( Zea mays L. cv. Northern Belle) seedlings chilled at 1.5°C in the dark for 3–30 h were compared with the ability of plants to resume growth in the immediate post-chilling period and with the development of visible symptoms of injury to the leaves. During chilling, the maximal rate of increase of the induced chlorophyll fluorescence rise. F_R, was measured on secondary leaf tissue. F_R decreased exponentially, at approximately the same rate in plants grown and chilled in hydroponic pots, in leaves detached from similar plants and in plants that were removed from the hydroponic pots and laid on wet filter paper adjacent to the detached leaves. The half-fall time for F_R in the 3 treatments was 7.8 ± 1.3 h, 8.6 ± 0.6 h and 8.8 ± 1.0 h, respectively. Following seedling removal from 1.5°C and return to 25/15°C, relative growth rates were determined from daily measurements of plant fresh weight gain. Compared with non-chilled seedlings, plants chilled for 3 h and longer showed depressed rates of growth. Inhibition of growth in the immediate post-chilling period (0–27 h) was linearly related to the duration of the chilling period and had a high positive correlation with the decrease in chlorophyll fluorescence (linearly related to log F_R) sustained during the chilling exposure. Visible symptoms of chilling injury developed during the post-chilling period on seedlings chilled for longer than 3 h. The decrease in log F_R during chilling was also linearly correlated with the severity of visual symptoms of chilling injury expressed in the post-chilling period. It is concluded that the extent of chilling injury in maize can be rapidly and non-destructively assessed from measurements of chlorophyll fluorescence.     

达乌尔黄鼠心肌细胞膜电位的耐寒性

用胞内微电极记录达乌尔黄鼠心室乳头肌细胞膜电位,研究它的耐寒性并附带观察季节及冬眠的影响。76％和55％的心肌标本分别在0℃至—5℃之间可诱发动作电位。3例心肌超冷至-5℃时静息电位(RP)66.6mV,为35℃时的80％,动作电位幅值(APA)60.4mV,为66％,但复温后可完全恢复。RP与APA比最大去极化率(dV/dt_(max))和动作电位时程(ADP)对寒冷有较大的抵抗力。季节对心肌膜电位活动有明显的影响,冬季深眠黄鼠的心肌RP和APA在0℃中显著高于冬季活跃组,提示深眠黄鼠的心肌有较大的耐寒性。这些结果说明冬眠型哺乳动物的心肌细胞膜电位比非冬眠型的有显著较大耐寒性。     

Helium‐based cold atmospheric plasma‐induced reactive oxygen species‐mediated apoptotic pathway attenuated by platinum nanoparticles

Plasma is generated by ionizing gas molecules. Helium (He)‐based cold atmospheric plasma (CAP) was generated using a high‐voltage power supply with low‐frequency excitation (60 Hz at 7 kV) and He flow at 2 l/min. Platinum nanoparticles (Pt‐NPs) are potent antioxidants due to their unique ability to scavenge superoxides and peroxides. These features make them useful for the protection against oxidative stress‐associated pathologies. Here, the effects of Pt‐NPs on He‐CAP‐induced apoptosis and the underlying mechanism were examined in human lymphoma U937 cells. Apoptosis was measured after cells were exposed to He‐CAP in the presence or absence of Pt‐NPs. The effects of combined treatment were determined by observing the changes in intracellular reactive oxygen species (ROS) and both mitochondrial and Fas dependent pathway. The results indicate that Pt‐NPs substantially scavenge He‐CAP‐induced superoxides and peroxides and inhibit all the pathways involved in apoptosis execution. This might be because of the SOD/catalase mimetic effects of Pt‐NPs. These results showed that the Pt‐NPs can induce He‐CAP desensitization in human lymphoma U937 cells.